Belgian Pig Research Articles

One Health surveillance of colistin-resistant Enterobacterales in Belgium and the Netherlands between 2017 and 2019.

Colistin serves as the last line of defense against multidrug resistant Gram-negative bacterial infections in both human and veterinary medicine. This study aimed to investigate the occurrence and spread of colistin-resistant Enterobacterales (ColR-E) using a One Health approach in Belgium and in the Netherlands. In a transnational research project, a total of 998 hospitalized patients, 1430 long-term care facility (LTCF) residents, 947 children attending day care centres, 1597 pigs and 1691 broilers were sampled for the presence of ColR-E in 2017 and 2018, followed by a second round twelve months later for hospitalized patients and animals. Colistin treatment incidence in livestock farms was used to determine the association between colistin use and resistance. Selective cultures and colistin minimum inhibitory concentrations (MIC) were employed to identify ColR-E. A combination of short-read and long-read sequencing was utilized to investigate the molecular characteristics of 562 colistin-resistant isolates. Core genome multi-locus sequence typing (cgMLST) was applied to examine potential transmission events. The presence of ColR-E was observed in all One Health sectors. In Dutch hospitalized patients, ColR-E proportions (11.3 and 11.8% in both measurements) were higher than in Belgian patients (4.4 and 7.9% in both measurements), while the occurrence of ColR-E in Belgian LTCF residents (10.2%) and children in day care centres (17.6%) was higher than in their Dutch counterparts (5.6% and 12.8%, respectively). Colistin use in pig farms was associated with the occurrence of colistin resistance. The percentage of pigs carrying ColR-E was 21.8 and 23.3% in Belgium and 14.6% and 8.9% in the Netherlands during both measurements. The proportion of broilers carrying ColR-E in the Netherlands (5.3 and 1.5%) was higher compared to Belgium (1.5 and 0.7%) in both measurements. mcr-harboring E. coli were detected in 17.4% (31/178) of the screened pigs from 7 Belgian pig farms. Concurrently, four human-related Enterobacter spp. isolates harbored mcr-9.1 and mcr-10 genes. The majority of colistin-resistant isolates (419/473, 88.6% E. coli; 126/166, 75.9% Klebsiella spp.; 50/75, 66.7% Enterobacter spp.) were susceptible to the critically important antibiotics (extended-spectrum cephalosporins, fluoroquinolones, carbapenems and aminoglycosides). Chromosomal colistin resistance mutations have been identified in globally prevalent high-risk clonal lineages, including E. coli ST131 (n = 17) and ST1193 (n = 4). Clonally related isolates were detected in different patients, healthy individuals and livestock animals of the same site suggesting local transmission. Clonal clustering of E. coli ST10 and K. pneumoniae ST45 was identified in different sites from both countries suggesting that these clones have the potential to spread colistin resistance through the human population or were acquired by exposure to a common (food) source. In pig farms, the continuous circulation of related isolates was observed over time. Inter-host transmission between humans and livestock animals was not detected. The findings of this study contribute to a broader understanding of ColR-E prevalence and the possible pathways of transmission, offering insights valuable to both academic research and public health policy development.

Genetic characterization of ESBL-producing and ciprofloxacin-resistant Escherichia coli from Belgian broilers and pigs.

The increasing number of infections caused by Escherichia coli resistant to clinically important antibiotics is a global concern for human and animal health. High overall levels of extended-spectrum beta-lactamase (ESBL)-producing and ciprofloxacin-resistant (ciproR) Escherichia coli in livestock are reported in Belgium. This cross-sectional study aimed to genotypically characterize and trace ESBL-and ciproR-E. coli of Belgian food-producing animals. A total of 798 fecal samples were collected in a stratified-random sampling design from Belgian broilers and sows. Consequently, 77 ESBL-E. coli and 84 ciproR-E. coli were sequenced using Illumina MiSeq. Minimum inhibitory concentration (MIC) for fluoroquinolones and cephalosporins were determined. Molecular in silico typing, resistance and virulence gene determination, and plasmid identification was performed. Scaffolds harboring ESBL or plasmid-mediated quinolone resistance (PMQR) genes were analyzed to detect mobile genetic elements (MGEs) and plasmid origins. Core genome allelic distances were used to determine genetic relationships among isolates. A variety of E. coli sequence types (ST) (n = 63), resistance genes and virulence profiles was detected. ST10 was the most frequently encountered ST (8.1%, n = 13). The pandemic multidrug-resistant clone ST131 was not detected. Most farms harbored more than one ESBL type, with bla CTX-M-1 (41.6% of ESBL-E. coli) being the most prevalent and bla CTX M-15 (n = 3) being the least prevalent. PMQR genes (15.5%, n = 13) played a limited role in the occurrence of ciproR-E. coli. More importantly, sequential acquisition of mutations in quinolone resistance-determining regions (QRDR) of gyrA and parC led to increasing MICs for fluoroquinolones. GyrA S83L, D87N and ParC S80I mutations were strongly associated with high-level fluoroquinolone resistance. Genetically related isolates identified within the farms or among different farms highlight transmission of resistant E. coli or the presence of a common reservoir. IncI1-I(alpha) replicon type plasmids carried different ESBL genes (bla CTX-M-1, bla CTX-M-32 and bla TEM-52C). In addition, the detection of plasmid replicons with associated insertion sequence (IS) elements and ESBL/PMQR genes in different farms and among several STs (e.g., IncI1-I(alpha)/IncX3) underline that plasmid transmission could be another important contributor to transmission of resistance in these farms. Our findings reveal a multifaceted narrative of transmission pathways. These findings could be relevant in understanding and battling the problem of antibiotic resistance in farms.

A Randomized Large-Scale Cross-Sectional Serological Survey of Hepatitis E Virus Infection in Belgian Pig Farms.

Hepatitis E virus (HEV) is the causative agent of hepatitis E disease in humans. While sporadic HEV infections, which occur in industrialised countries and are typically due to HEV genotypes 3 or 4, are asymptomatic and self-limiting, a chronic form of the disease can lead to liver cirrhosis in immunocompromised individuals. Pigs share HEV 3 and 4 genotypes and are thus considered a major animal reservoir for human infection. A subset of animals has been shown to carry HEV particles at the age of slaughter, rendering raw or undercooked pig products potential vectors for human infection. To provide an overview of the current dissemination of HEV in Belgian pig herds, this study was designed as a randomized, robust, large-scale, cross-sectional, serological survey. HEV genotypes and subtypes recently circulating in Belgium (2020-2021) were investigated. Sample stratification as well as epidemiological investigation through the available demographic data of the sampled herds showed that HEV widely circulated in the Belgian pig population during this time and that a change in the circulating HEV strains may have occurred in the last decade. Herd size and type were identified as risk factors for HEV herd-seropositivity. Identifying farms at risk of being HEV-positive is an important step in controlling HEV spread and human infection.

Porcine ear necrosis in weaned piglets: prevalence and impact on daily weight gain

BackgroundPorcine ear necrosis (PEN) in pigs is characterized by a blue to black discoloration of the tip or margin of the ear followed by necrosis. The present study investigated the prevalence of PEN in a Belgian pig farm with PEN problems in nursery pigs, the effect of a mycotoxin detoxifier added to the feed on PEN prevalence, and the impact of PEN on the piglets’ growth. Six consecutive batches of weaned piglets [565–751 piglets per batch, (n = 3898)] were included. For each weaning batch, the presence and severity of PEN during the nursery period (3–10 weeks of age) were recorded weekly. Average daily gain (ADG) was calculated by weighing 597 individual piglets divided over the six batches. Additionally different mycotoxins were measured in the feed using LC–MS/MS analysis, and to three randomly selected batches, a mycotoxin detoxifier (Mycofix® Plus 5E, Biomin) was added to the feed.ResultsAt the end of the nursery period, 11.0–32.0% of the piglets in each batch were affected. The prevalence increased with the number of weeks post-weaning, especially from week 4 after weaning onwards. Mild, moderate, severe and very severe lesions represented 84.6%, 14.0%, 1.3% and 0.1% of all lesions, respectively. Different mycotoxins were present in the feed, but all at low concentrations. The mean ADG (± SD) for pigs without (n = 243) and with (n = 158) lesions was 391 g (± 71 g) and 394 g (± 65 g), respectively (P > 0.05). The ADG for mildly affected (387 g ± 68 g) and moderately affected piglets (420 g ± 44 g) was not significantly different (P > 0.05). The PEN prevalence in the batches without or with the mycotoxin detoxifier was 25% and 22%, respectively (P > 0.05).ConclusionsTwenty-three percent of animals showed lesions at the end of the nursery. Affected pigs did not have a lower ADG compared to non-affected animals, which might be explained by the fact that most affected piglets only had mild lesions. The addition of a mycotoxin detoxifier did not influence the prevalence of PEN, possibly because of the low levels of mycotoxin contamination. Further research is warranted to assess the impact of more severe PEN lesions and the effect of control measures.

Seasonal Variation in Prevalence of Mycoplasma hyopneumoniae and Other Respiratory Pathogens in Peri-Weaned, Post-Weaned, and Fattening Pigs with Clinical Signs of Respiratory Diseases in Belgian and Dutch Pig Herds, Using a Tracheobronchial Swab Sampling Technique, and Their Associations with Local Weather Conditions.

Besides Mycoplasma hyopneumoniae (M. hyopneumoniae), many other viruses and bacteria can concurrently be present in pigs. These pathogens can provoke clinical signs, known as porcine respiratory disease complex (PRDC). A sampling technique on live animals, namely tracheobronchial swab (TBS) sampling, was applied to detect different PRDC pathogens in pigs using PCR. The objective was to determine prevalence of different PRDC pathogens and their variations during different seasons, including correlations with local weather conditions. A total of 974 pig farms and 22,266 pigs were sampled using TBS over a 5-year period. TBS samples were analyzed using mPCR and results were categorized and analyzed according to the season of sampling and local weather data. In samples of peri-weaned and post-weaned piglets, influenza A virus in swine (IAV-S), porcine reproductive and respiratory syndrome virus—European strain (PRRSV1), and M. hyopneumoniae were found as predominant pathogens. In fattening pigs, M. hyopneumoniae, porcine circovirus type 2 (PCV-2) and PRRSV1 were predominant pathogens. Pathogen prevalence in post-weaned and finishing pigs was highest during winter, except for IAV-S and A. pleuropneumoniae, which were more prevalent during autumn. Associations between prevalence of several PRDC pathogens, i.e., M. hyopneumoniae, PCV-2 and PRRSV, and specific weather conditions could be demonstrated. In conclusion, the present study showed that many respiratory pathogens are present during the peri-weaning, post-weaning, and fattening periods, which may complicate the clinical picture of respiratory diseases. Interactions between PRDC pathogens and local weather conditions over the 5-year study period were demonstrated.

ESBL-Producing, Carbapenem- and Ciprofloxacin-Resistant Escherichia coli in Belgian and Dutch Broiler and Pig Farms: A Cross-Sectional and Cross-Border Study.

Background. The use of antibiotics in food production selects for resistant bacteria and may cause a threat to human and animal health. Belgium and the Netherlands have one of the highest densities of broilers and pigs in Europe, making active monitoring of antibiotic use and resistance in this region vital. Objectives. This study aimed to quantify ESBL-producing (ESBL-E. coli), carbapenem- and ciprofloxacin-resistant (CiproR) Escherichia coli in animal feces on broiler and pig farms with a history of high antibiotic use in Belgium and the Netherlands. Methods. A total of 779 broiler and 817 pig fecal samples, collected from 29 conventional broiler and 31 multiplier pig farms in the cross-border region of Belgium and the Netherlands, were screened for the presence of antibiotic-resistant E. coli using selective culturing. Results. Carbapenem-resistant E. coli were not detected. ESBL-E. coli were remarkably more prevalent in samples from Belgian than Dutch farms. However, CiproR-E. coli were highly prevalent in broilers of both countries. The percentage of samples with ESBL- and CiproR-E. coli was lower in pig compared to poultry farms and varied between farms. No clear association with the on-farm antibiotic use in the year preceding sampling was observed. Multidrug resistance was frequently observed in samples from both countries, but ESBL-production in combination with ciprofloxacin resistance was higher in samples from Belgium. Conclusions. This study demonstrated marked differences in antibiotic resistance between countries, farms and within farms. The observed variation cannot be explained straightforward by prior quantity of antibiotic use suggesting that it results from more complex interactions that warrant further investigation.

Genetic and antigenic evolution of H1 swine influenza A viruses isolated in Belgium and the Netherlands from 2014 through 2019

Surveillance of swine influenza A viruses (swIAV) allows timely detection and identification of new variants with potential zoonotic risks. In this study, we aimed to identify swIAV subtypes that circulated in pigs in Belgium and the Netherlands between 2014 and 2019, and characterize their genetic and antigenic evolution. We subtyped all isolates and analyzed hemagglutinin sequences and hemagglutination inhibition assay data for H1 swIAV, which were the dominant HA subtype. We also analyzed whole genome sequences (WGS) of selected isolates. Out of 200 samples, 89 tested positive for swIAV. swIAV of H1N1, H1N2 and H3N2 subtypes were detected. Analysis of WGS of 18 H1 swIAV isolates revealed three newly emerged genotypes. The European avian-like H1 swIAV (lineage 1C) were predominant and accounted for 47.2% of the total isolates. They were shown to evolve faster than the European human-like H1 (1B lineage) swIAV, which represented 27% of the isolates. The 2009 pandemic H1 swIAV (lineage 1A) accounted for only 5.6% of the isolates and showed divergence from their precursor virus. These results point to the increasing divergence of swIAV and stress the need for continuous surveillance of swIAV.

Doing the “dirty work”: how hunters were enlisted in sanitary rituals and wild boars destruction to fight Belgium's ASF (African Swine Fever) outbreak

In the early autumn of 2018, a virus as contagious as it is deadly, carried by wild boars (Sus scrofa Linnaeus, 1758) with the probable involvement of humans, crossed the Belgian border. African swine fever, which only affects suidae, is rapidly spreading in the forests of Gauma. The boar, whose status has gradually shifted from a regional emblem to a symbol of hunting abuses, finds itself abruptly transformed into a sanitary threat needing to be eliminated. The wild swine can contaminate its domestic cousin, the farmed pig (Sus domesticus Erxleben, 1777). Therefore, the spread of the virus would jeopardise the fragile Belgian pig farming sector concentrated in the north of the country. This is the start of a crisis that will last for more than 24 months; the infected forest is zoned and then isolated for the purpose of sanitisation, while “biosecurity” and “white zone” become the only watchwords. Mass destruction measures for wild boars are imposed by the administration and its experts through new so-called “sanitary rituals”. To achieve a rapid “return to normal”, hunters – mostly local ones – are enlisted in the name of their hunting skills, which, although they are usually contested by a part of Belgian society and media, are considered essential in this case. This event brings us to an exploration of the practices actors are attached to and forced to renounce to in the name of good crisis management. On-the-ground realities as related by field men bear witness to the unease felt in the face of the “dirty work” asked of them, while the upheaval of coexistence reveals ethical, tradition- and identity-related questions already existing before the crisis.

Purchasing policy, quarantine and acclimation practices of breeding gilts in Belgian pig farms

BackgroundThe breeding population is very important in pig herds, for productivity, health and profitability. Replacement of breeding animals can be accomplished by own rearing of breeding gilts or by purchasing them. Purchasing breeding gilts is a hazardous event in terms of biosecurity and introduction of pathogens into a farm. However, in literature, little is known about gilt introduction in a herd. The present study investigated the introduction procedures of purchased breeding gilts in Belgian pig herds, and the compliance of these herds to the optimal introduction procedures. A questionnaire consisting of twenty questions related to farm characteristics (n = 2), purchasing policy (n = 6), quarantine period (n = 5), and acclimation practices (n = 7) was designed, and 68 farms completed the questionnaire during an on-farm interview.ResultsThe median (min. – max.) number of sows on the farms was 300 (85–2500). Fifty-seven per cent of the farms purchased breeding gilts, and there was a lot of variation in the frequency of purchase and the age at which gilts are purchased. On 95 % of those farms, a quarantine unit was used, and on most of these farms the quarantine was located on the farm itself (internal quarantine). The median (min. – max.) duration of the quarantine period was 42 (14–140) days. The most common acclimation practice was vaccination against Porcine parvovirus (96 %) and Erysipelothrix rhusiopathiae (94 %), although in some farms exposure of gilts to farm-specific micro-organisms was done by providing faeces from suckling piglets (18 %) and bringing gilts in contact with sows that will be culled (16 %). Only 10 % of the farms complied with the optimal introduction procedures, i.e. purchasing policy, quarantine building and quarantine management.ConclusionsThis study showed that in many farms, practices related to purchasing, quarantine and acclimation could be improved to maintain optimal biosecurity.

Sources of variation of antimicrobial use in Charolaise and Limousine beef breeds in Veneto region (Italy)

Sources of variation of antimicrobial use in Charolaise and Limousine beef breeds in Veneto region (Italy)

Combining quantitative and qualitative approaches to determine viability of a potential Salmonella Typhimurium vaccination program in pigs in Belgium

Vaccinating pigs against Salmonella Typhimurium (ST) might be a way to control ST infections at farm level and reduce human infections. Two main issues have to be addressed before such a mandatory vaccination program can be implemented: the effective reduction of attributable human incidence has to be demonstrated and all socio-economic barriers impacting the attitude and motivation of the pig sector have to be lifted.The present research used a quantitative microbial risk assessment model to estimate the effect of different hypothetical Salmonella spp. and ST mitigation strategies on the annual prevalence of human salmonellosis along the minced pork production chain. In addition, a qualitative study aimed to list the potential concerns of the pig sector about the implementation of a hypothetical future vaccination program.The following themes were the most often mentioned: awareness, vaccine cost-benefit/effectiveness, legislation, monovalent vaccine, time and labour required to vaccinate, vaccine registration and trade restriction. Cost-effectiveness and cost-benefit of vaccination were cited by all the key interviewees (n = 12). However, based on the quantitative microbial risk assessment model, vaccination alone may not be sufficiently effective to reduce the annual human salmonellosis prevalence. A combination of different control measures along the food chain, with a special focus on interventions at the slaughterhouse, might be more effective in achieving the desired goal than vaccination alone.

Stability, Homogeneity and Carry-Over of Amoxicillin, Doxycycline, Florfenicol and Flubendazole in Medicated Feed and Drinking Water on 24 Pig Farms.

The vast majority of medicines in pig rearing are administered via oral group medication through medicated feed and drinking water. However, relevant on-farm factors affecting the concentration of these drugs in feed and drinking water, such as the homogeneity, stability, and cross-contamination, are largely unknown. To characterize these factors, samples of medicated feed and drinking water were taken on 24 Belgian pig farms during treatment and 2 days thereafter, as well as at different on-farm sampling sites from production to feeding troughs or drinking nipples. The samples contained amoxicillin, doxycycline, florfenicol, or flubendazole. Additionally, a questionnaire was completed. In contrast to the results of medicated feed, results of medicated water showed a large between-farm variation in antimicrobial drug concentration. The therapeutic concentration range was only met in 2 out of 11 farms using medicated feed, and in 3 out of 13 farms using medicated water. Medicated feed concentrations were often below the therapeutic concentration range mentioned in the Summary of Product Characteristics, while drinking water concentrations were just as often above as they were below the advised target concentration range. Drug residues measured 2 days after the end of therapy with both feed and water medication rarely exceeded 1% of the lowest therapeutic concentration. This study demonstrates that recommendations on good clinical practices for oral group medication in the pig industry are highly needed.

Can oral group medication be improved to reduce antimicrobial use?

Of the antimicrobials sold to treat food-producing animals in Europe in 2016, 90.1 per cent were designated for group treatment.1 Group treatment, administered through drinking water or feed, enables farmers to easily treat a large number of animals as soon as clinical signs of disease are observed in the group. This practice – referred to as metaphylaxis or control treatment – has the advantage of quickly treating many animals at the same time in order to slow the spread of disease. It has been shown that metaphylactic treatments are associated with higher cure rates than curative treatments administered when animals are already exhibiting clinical signs of disease.2,3 However, the European guidelines for the prudent use of antimicrobials in veterinary medicine4 highlight treatment via feed and drinking water as a major concern in the development of antimicrobial resistance. To help limit the emergence and spread of antimicrobial resistance, it is necessary to control the concentration of antimicrobials in feed and drinking water, the quantity delivered to the animals and the quantity ingested by each animal. In a study summarised on p 405 of this issue of Vet Record , Vandael and colleagues investigate the use of medicated feed and drinking water on Belgian pig farms and discuss the hurdles that have to be overcome to meet these three criteria.5 Vandael and colleagues found discrepancies in the preparation of medicated feed and drinking water. The concentration incorporated into feed seems to be reasonably well controlled, with 68.2 per cent of the pig farms surveyed purchased premixed medicated feed from a manufacturer. However, the use of …

Oral group medication in pig production: characterising medicated feed and drinking water systems.

Despite common use of oral group medication in pig rearing, the homogeneity, stability and carry-over of frequently used medicinal products in feed and drinking water are largely unknown. Therefore, a field study was performed on 52 Belgian pig farms, characterising preparation and administration of medicinal products via these systems, and farmers' user experiences with medicated feed and medicated drinking water. The study showed that medicated drinking water is more commonly used than medicated feed, since 90.4 per cent of the farms sometimes use medicated drinking water and 69.2 per cent of the farms sometimes use medicated feed. The drinking water quality is evaluated at least once a year on only 30.7 per cent of the farms. Separate pipelines for medicated and non-medicated circuits were not present in any of the farms using medicated feed and in 27.7 per cent of the farms using medicated drinking water. With drinking water medication, 63.5 per cent of the farmers reported encountering practical problems, often related to solubility issues and precipitation of the active compounds. In contrast, medicated feed is bought ready-to-use from the feed manufacturer in 68.2 per cent of the cases, thus reducing the number of practical problems experienced by the farmer. This study shows room for improvement of oral group treatment, developing appropriate pharmaceutical formulations for drinking water medication, quality control of drinking water, using separate pipeline circuits, and cleaning and disinfecting protocols.

Prevalence and chemical composition of uroliths in fattening pigs in Belgium

The present study investigated the prevalence of uroliths in fattening pigs and assessed the composition of these urinary tract concrements. In total, 2,432 urinary bladders were sampled in the slaughterhouse and checked for abnormal content. Urinary samples were analysed microscopically, and samples of the urinary bladder wall were tested for histological signs of inflammation. The composition of the concrements was examined by infrared spectrophotometry. Macroscopic and microscopic abnormalities were detected in 8.4% and 52.8% of the samples respectively. Magnesium ammonium phosphate (struvite), calcium oxalate dihydrate (COD), calcium carbonate (calcite), calcium oxalate monohydrate (COM) and amorphous crystals were detected. Analysis of stones showed COD in all samples in different proportions. The calcium content of examined stones was always considerable (up to 34%), in contrast to the magnesium content which represented max 1.9%. Struvite was found in one third of the samples, but was never part of stones and grit. COD crystals were the second most common microscopic crystal. These COD crystals and some COD stones had a rectangular shape, and therefore, they can be harmful to the bladder mucosa. In conclusion, uroliths are present in a large proportion of male fattening pigs, and consequently, urinary concrements pose a life-threatening risk for urethra obstruction in male pigs. Further research is warranted to identify potential risk factors for urolithiasis and microscopic crystals.

Development and validation of an LC–MS/MS method for the simultaneous determination of citrinin and ochratoxin a in a variety of feed and foodstuffs

An ultra-performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-ESI+/−-MS/MS) method for the simultaneous analysis of citrinin (CIT) and ochratoxin A (OTA) in feed (chicken and pig) and food (cereal-based products, fruit, vegetable juices, nuts, seeds, herbs, spices, vegetarian and soy products, alcoholic beverages, baby food products and food supplements) was developed. The mycotoxins were extracted from these matrices using a QuEChERS-based extraction method without any further clean-up step. The samples were 5-fold concentrated. Final extracts were analyzed using a UPLC–MS/MS system and chromatographic separation was achieved by applying a gradient elution for a total run time of 10 min. Mycotoxins were quantified using an internal calibration via analyte/13C-labeled internal standard ratio. The developed method was validated according to the criteria described in Commission Regulation No. 401/2006/EC and Commission Decision No. 2002/657/EC. Specificity, linearity, apparent recovery, limit of detection and quantification, intraday and interday precision, measurement uncertainty, matrix effect, and extraction efficiency were the parameters studied. Finally, 90 Belgian chicken and pig feed samples were analyzed, revealing the simultaneous presence of CIT (<LOQ − 3.90 μg/kg) and OTA (<LOQ − 5.60 μg/kg) in more than 50% of these products.

The effect on early renal function of various dynamic preservation strategies in a preclinical pig ischemia-reperfusion autotransplant model.

The aims of this study were to determine the most optimal timing to start machine perfusion during kidney preservation to improve early graft function and to evaluate the impact of temperature and oxygen supply during machine perfusion in a porcine ischemia-reperfusion autotransplant model. The left kidney of an approximately 40-kg female Belgian Landrace pig was exposed to 30 minutes of warm ischemia via vascular clamping and randomized to 1 of 6 study groups: (1) 22-hour static cold storage (SCS) (n = 6), (2) 22-hour hypothermic machine perfusion (HMP) (n = 6), (3) 22-hour oxygenated HMP (n = 7), (4) 20-hour HMP plus 2-hour normothermic perfusion (NP) (n = 6), (5) 20-hour SCS plus 2-hour oxygenated HMP (n = 7), and (6) 20-hour SCS plus 2-hour NP (n = 6). Graft recovery measured by serum creatinine level was significantly faster for continuous HMP preservation strategies compared with SCS alone and for all end-ischemic strategies. The active oxygenated 22-hour HMP group demonstrated a significantly faster recovery from early graft function compared with the 22-hour nonactive oxygenated HMP group. Active oxygenation was also found to be an important modulator of a faster increase in renal flow during HMP preservation. Continuous oxygenated HMP applied from the time of kidney procurement until transplant might be the best preservation strategy to improve early graft function.

Correlation between slaughter practices and the distribution of Salmonella and hygiene indicator bacteria on pig carcasses during slaughter

This study investigated the distribution of hygiene indicator bacteria and Salmonella on pig carcasses. Moreover, the relation between hygiene indicator counts and Salmonella presence as well as associations between specific slaughter practices and carcass contamination were determined for each carcass area. Seven Belgian pig slaughterhouses were visited three times to swab five randomly selected carcasses at nine different areas, after evisceration and trimming. Information about slaughter practices was collected using a questionaire. In all samples, the E. coli and Salmonella presence was analyzed and Enterobacteriaceae and total aerobic bacteria were quantified. Average total aerobic counts ranged from 3.1 (loin, pelvic duct, ham) to 4.4 log10 CFU/cm2 (foreleg). Median Enterobacteriaceae numbers varied between 0.4 (ham) an 1.8 log10 CFU/cm2 (foreleg). E. coli and Salmonella presence ranged from 15% (elbow) to 89% (foreleg) and 5% (elbow) to 38% (foreleg), respectively. Positive relations were found between hygiene indicator counts and Salmonella presence at the head, sternum, loin and throat. Several slaughter practices, such as splitting the head and incising tonsils, were associated with higher levels of hygiene indicator bacteria and Salmonella. These findings can be used to educate slaughterhouse personnel and estimate the public health risk involved in consumption of different pork cuts.

Sequence-Based Genotyping of Expressed Swine Leukocyte Antigen Class I Alleles by Next-Generation Sequencing Reveal Novel Swine Leukocyte Antigen Class I Haplotypes and Alleles in Belgian, Danish, and Kenyan Fattening Pigs and Göttingen Minipigs.

The need for typing of the swine leukocyte antigen (SLA) is increasing with the expanded use of pigs as models for human diseases and organ-transplantation experiments, their use in infection studies, and for design of veterinary vaccines. Knowledge of SLA sequences is furthermore a prerequisite for the prediction of epitope binding in pigs. The low number of known SLA class I alleles and the limited knowledge of their prevalence in different pig breeds emphasizes the need for efficient SLA typing methods. This study utilizes an SLA class I-typing method based on next-generation sequencing of barcoded PCR amplicons. The amplicons were generated with universal primers and predicted to resolve 68–88% of all known SLA class I alleles dependent on amplicon size. We analyzed the SLA profiles of 72 pigs from four different pig populations; Göttingen minipigs and Belgian, Kenyan, and Danish fattening pigs. We identified 67 alleles, nine previously described haplotypes and 15 novel haplotypes. The highest variation in SLA class I profiles was observed in the Danish pigs and the lowest among the Göttingen minipig population, which also have the highest percentage of homozygote individuals. Highlighting the fact that there are still numerous unknown SLA class I alleles to be discovered, a total of 12 novel SLA class I alleles were identified. Overall, we present new information about known and novel alleles and haplotypes and their prevalence in the tested pig populations.

Characterization of porcine endogenous retrovirus expression in neonatal and adult pig pancreatic islets.

Pig islets represent an alternative to the current modes of treatment for patients with diabetes. However, the concerns over pathogen transmission including that of PERV limit their immediate, widespread usage in humans. It has been previously demonstrated that PERV copy number and particularly expression levels can vary considerably between individuals and within different tissues of a single animal. In general, expression levels have been found to be particularly low in the pancreas compared to other porcine tissues suggesting a reduced risk associated with the use of this tissue. Data regarding this crucial aspect, however, remain limited and little is known about PERV status of islets themselves, which represent the final product to be transplanted. In addition, comparative analysis of the PERV status of neonatal piglets with adults is important as they are increasingly considered as potential islet donors for xenotransplantation. Tissue samples from 51 neonatal piglets (age between 14 and 21days) and 29 adult pigs were collected from Belgian landrace pigs used for pancreas procurement and islet isolation. Tissue biopsies were used to extract DNA for PERV copy number quantification by qPCR and RNA for PERV expression by qRT-PCR. As expected, PERV expression demonstrated great variation and was significantly lower in pancreas compared to other tissues. More importantly, PERV RNA expression was found to be specifically enriched in pancreatic islets reaching values similar to those found in other tissues such as liver and kidney. Interestingly, this expression was not coupled with the detection of reverse transcriptase in islet cultures or indeed detection of PERV virus. Lung, spleen, and lymph node consistently showed the highest levels of PERV expression. Comparison of PERV in neonatal and adult pigs showed that copy number did not vary significantly from birth to adulthood. PERV expression on the other hand was significantly lower in neonatal pig islets compared to adult islets and did not increase over the period of culture. Our study confirms the low level of PERV expression in whole pancreas in a large population of both neonatal and adult pigs (n=80). The level of PERV expression was however higher in the endocrine tissue than in the exocrine cells. There was no correlation between PERV status in donor PBMCs and islet cells, and no evidence of active replication in in vitro regardless of PERV expression in islet cells. Moreover, neonatal pig islets were found to have significantly lower PERV expression compared to adult islets. Neonatal islets have been suggested as the best choice for xenotransplantation in terms of economic and procurement considerations; the PERV status reported here would also potentially support their use.